We have established and characterized a bovine SV40-Large-T-transduced cell line. For information on how many population doublings the cell line does until crisis, I want to calculate it.
Is it OK to use the population doubling time obtained from the logarithmic phase of a growth curve to calculate the population doublings or do I need to use the cell counts from passage 1 upwards in order to generate a graph showing the population doublings as done in many cell line characterization papers?
to my understanding you can only use PDs of the logarithmic phase this is defined in the mathematical formula as well:
3,32*(Log(cells at day x)-(cellas at day y)/Log(2)
That depends what you really want to know. Is it the population doblings of the whole population or is it the number of cell divisions. These two things are not equal if not all cells in your culture are actually cycling (as often is the case). To check, you may want to incubate your cells with BrdU (or EdU) for a time longer than it takes to double the population. If you then stain for the BrdU (or EdU), all cells cycling cells should have incorporated it, while all quiescent cells are negative. The fraction of dividing cells you can the use as a coefficient to calculate the real doubling time of the dividing cells ( which would be shorter than the population doubling time).
I also would prefer the BrdU-assay using different incubation times. For a "quick and dirty method" you can seed the cells in 3-5 wells on a 24 well plate (same cell number) and count them after different time point (for example after 24, 30 and 48h). Then you can use the formula of Dennis.
We already did a growth curve with a cell counter (Moxi Z) with good results.
Now I want to calculate the population doublings (PDL) as I want to get an information on how many PDLs our newly established cell line does until it starts to reach crisis with phenotypical changes.
Is is recommend to to this from the logarithmic phase of the data from the growth curve to obtain an estimate? From this data I plan to generate a graph (number of passages on x-axis, PDL on y-axis as seen in some cell characterization publications to visualize the data. Would this be OK?
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